RNA干扰Rac1基因表达对结直肠癌LoVo细胞骨架和细胞周期的影响  

Silencing Rac1gene expression via RNA interference affects cytoskeleton and cell cycle of colorectal cancer LoVo cells

作　　者：赵世义[1] 徐胜平[2] 李桂云[1] 李晓东[1] 栾祖鹏[1] 黎晓鹃[1] 

机构地区：[1]济南市第三人民医院肿瘤科,山东济南250101 [2]济南市疾病预防控制中心免疫预防所,山东济南250001

出　　处：《中国肿瘤生物治疗杂志》2012年第5期486-490,共5页Chinese Journal of Cancer Biotherapy

基　　金：济南市科技局基金资助项目(No.2007-153);济南市卫生局基金资助项目(No.2008-089)~~

摘　　要：目的:观察Rac1蛋白在结直肠癌细胞中的表达,并分析其与结直肠癌LoVo细胞骨架、细胞周期和细胞凋亡的相关性。方法:Western blotting测定5种结直肠癌细胞株(LoVo,SW480,SW620,SW1116,HT29)中Rac1蛋白的表达;Rac1-shR-NA质粒转染LoVo细胞后,激光共聚焦显微镜观察LoVo细胞骨架的变化,流式细胞仪检测细胞周期和细胞凋亡的变化。结果:5种结直肠癌细胞株中均有Rac1蛋白的高表达。Rac1基因沉默后,LoVo细胞中交联状态的F-actin网明显减少且紊乱,G0/G1期细胞比例较对照组显著增加[(74.63±4.40)%vs(56.46±3.09)%,P<0.05],而S期细胞比例较对照组显著减少[(12.87±1.77)%vs(29.66±1.92)%,P<0.05];Rac1-shRNA转染组LoVo细胞凋亡率较对照组显著增加[(25.31±2.05)%vs(9.38±1.16)%,P<0.05]。结论:RNA沉默Rac1基因的表达影响了LoVo细胞骨架的形成和细胞周期,为以Rac1为靶点治疗结直肠癌提供了实验依据。Objective：To detect the expression of Rac1 protein in colorectal cancer cells and to observe its correlation with cytoskeleton, cell cycle and apoptosis of colorectal cancer LoVo cells. Methods： The expression of Rac1 protein was detected by Western blotting in five colorectal cancer cell lines. The cytoskeleton changes were observed by confocal microscopy in LoVo cells transfected with Rac1-shRNA. The cell cycle and apoptosis were observed by flow cytometry in LoVo cells transfected with Rac1-shRNA. Results： Rac1 protein was highly expressed in five colorectal cancer cells. The cross-linked F-actin network was significantly reduced and disordered in LoVo cells transfected with Rac1-shRNA. The cell number in G0/G1 phase was significantly increased in Rac1-shRNA group compared with the control group （[74.63±440]% vs [56.46±3.09]%,P〈0.05）, whereas it was reduced in S phase （[12.87±1.77]% vs [29.66±192]%,P〈0.05） and the apoptosis rate of LoVo cells was significantly increased in Rac1-shRNA group compared with the control group （[25.31±2.05]% vs [9.38±1.16]%, P〈0.05）. Conclusion： Silencing Rac1 expression mediated by RNA interference affects the formation of cytoskeleton and the cell cycle of LoVo cells, which provides clinical evidence for Rac1-targeted treatment of colorectal cancer.

关 键 词：结直肠癌 RNA干扰 Rac1基因 细胞骨架 细胞周期 凋亡 

分 类 号：R735.3[医药卫生—肿瘤] R730.2[医药卫生—临床医学]